Abstract
BackgroundTigecycline is a last-resort antibiotic used to treat severe infections caused by extensively drug-resistant bacteria. Recently, novel tigecycline resistance genes tet(X3) and tet(X4) have been reported, which pose a great challenge to human health and food security. The current study aimed to establish a TaqMan-based real-time PCR assay for the rapid detection of the tigecycline-resistant genes tet(X3) and tet(X4).ResultsNo false-positive result was found, and the results of the TaqMan-based real-time PCR assay showed 100% concordance with the results of the sequencing analyses. This proposed method can detect the two genes at the level of 1 × 102 copies/μL, and the whole process is completed within an hour, allowing rapid screening of tet(X3) and tet(X4) genes in cultured bacteria, faeces, and soil samples.ConclusionTaken together, the TaqMan-based real-time PCR method established in this study is rapid, sensitive, specific, and is capable of detecting the two genes not only in bacteria, but also in environmental samples.
Highlights
Tigecycline is a last-resort antibiotic used to treat severe infections caused by extensively drugresistant bacteria
With the prevalence of antimicrobial resistance (AMR), only a few antibiotics are available to treat severe infections caused by extensively drug-resistant (XDR) bacteria, which poses a great challenge to human health and food security
Primers and probes The results of the National Center for Biotechnology Information (NCBI) Primer Basic Local Alignment Search Tool (BLAST) module indicated that no genes other than tet(X3) and tet(X4) genes matched the primer sequences designed in this study
Summary
Tigecycline is a last-resort antibiotic used to treat severe infections caused by extensively drugresistant bacteria. Novel tigecycline resistance genes tet(X3) and tet(X4) have been reported, which pose a great challenge to human health and food security. With the prevalence of antimicrobial resistance (AMR), only a few antibiotics are available to treat severe infections caused by extensively drug-resistant (XDR) bacteria, which poses a great challenge to human health and food security. Tet(X3) and tet(X4) are the first plasmid-borne tet(X) genes, encoding proteins with 386 amino acids and 385 amino acids, respectively, and showing 85.1 and 94.3% identity, respectively, with the original tet(X) from Bacterioides fragilis [4, 5] To date, both genes have been identified in humans, animals, meat, and environmental samples [4, 6,7,8]. In three representative provinces of China, tet(X3) and tet(X4) genes have been identified in animals and meat for consumption
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